Bihoreau N, Pin S, de Kersabiec A M, Vidot F, Fontaine-Aupart M P
TM Innovation (Centre National de Transfusion Sanguine Institut Mérieux, Les Ulis, France.
C R Acad Sci III. 1993;316(5):536-9.
Anti-hemophilia A factor (FVIII) consists in different heterodimers of heavy and light chains associated by a metallic ion. The integrity of this complex is indispensable for procoagulant activity. Atomic absorption spectrometry with Zeeman effect has been applied to determine the nature of this metal. For this purpose, the different active forms of FVIII were separated by FPLC and characterized by SDS-PAGE. Two peaks were observed, the first corresponding to different FVIII complexes of high molecular mass (ranging from 210-80 kDa to 110-80 kDa) and the second to the heavy-light chain dimer of 90-80 kDa. In all these active fractions, copper atom was identified and a proportionality was measured between the metal concentration and the coagulant activity. Furthermore, the determination of copper and FVIII concentrations indicated that only one copper atom is implicated in the 90-80 kDa association.
抗血友病因子A（FVIII）由通过金属离子结合的重链和轻链的不同异二聚体组成。该复合物的完整性对于促凝活性不可或缺。采用带塞曼效应的原子吸收光谱法来确定这种金属的性质。为此，通过快速蛋白质液相色谱法（FPLC）分离FVIII的不同活性形式，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）进行表征。观察到两个峰，第一个峰对应于不同的高分子量FVIII复合物（范围从210 - 80 kDa到110 - 80 kDa），第二个峰对应于90 - 80 kDa的重-轻链二聚体。在所有这些活性组分中，鉴定出了铜原子，并测定了金属浓度与凝血活性之间的比例关系。此外，铜和FVIII浓度的测定表明，在90 - 80 kDa的缔合物中仅涉及一个铜原子。
